LOCUS       sequence-183793-        7036 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7036)
  AUTHORS   Cho W, Hagemann TL, Johnson DA, Johnson JA, Messing A
  TITLE     Dual transgenic reporter mice as a tool for monitoring expression of
            glial fibrillary acidic protein.
  JOURNAL   J Neurochem. 2009 Jul;110(1):343-51. Epub 2009 May 5.
  PUBMED    19457099
REFERENCE   2  (bases 1 to 7036)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J 
            Neurochem."; date: "2009-07"; volume: "110(1)"; pages: "343-51. Epub
            2009 May 5"
COMMENT     Sequence Label: hGFAP-fLuc
FEATURES             Location/Qualifiers
     source          1..7036
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    7..55
                     /note="synthetic polyadenylation signal"
     misc_feature    69..160
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     109..128
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      2467..2476
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2473..4125
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(2539..2557)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    4166..4287
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4177..4196)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4231..4250
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4535..4552)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4706..5294)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4786..4805)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5465..6256,6257..6325))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6326..6430)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6457..6912
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6544..6563)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6754..6775
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg
       61 atagtactaa catacgctct ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag
      121 gctgtcccca gtgcaagtgc aggtgccaga acatttctct atcgataggt accgagctct
      181 tacgcgtgct agcccgggct cgagatctga gctcccacct ccctctctgt gctgggactc
      241 acagagggag acctcaggag gcagtctgtc catcacatgt ccaaatgcag agcataccct
      301 gggctgggcg cagtggcgca caactgtaat tccagcactt tgggaggctg atgtggaagg
      361 atcacttgag cccagaagtt ctagaccagc ctgggcaaca tggcaagacc ctatctctac
      421 aaaaaaagtt aaaaaatcag ccacgtgtgg tgacacacac ctgtagtccc agctattcag
      481 gaggctgagg tgaggggatc acttaaggct gggaggttga ggctgcagtg agtcgtggtt
      541 gcgccactgc actccagcct gggcaacagt gagaccctgt ctcaaaagac aaaaaaaaaa
      601 aaaaaaaaaa aaaagaacat atcctggtgt ggagtagggg acgctgctct gacagaggct
      661 cgggggcctg agctggctct gtgagctggg gaggaggcag acagccaggc cttgtctgca
      721 agcagacctg gcagcattgg gctggccgcc ccccagggcc tcctcttcat gcccagtgaa
      781 tgactcacct tggcacagac acaatgttcg gggtgggcac agtgcctgct tcccgccgca
      841 ccccagcccc cctcaaatgc cttccgagaa gcccattgag cagggggctt gcattgcacc
      901 ccagcctgac agcctggcat cttgggataa aagcagcaca gccccctagg ggctgccctt
      961 gctgtgtggc gccaccggcg gtggagaaca aggctctatt cagcctgtgc ccaggaaagg
     1021 ggatcagggg atgcccaggc atggacagtg ggtggcaggg ggggagagga gggctgtctg
     1081 cttcccagaa gtccaaggac acaaatgggt gaggggactg ggcagggttc tgaccctgtg
     1141 ggaccagagt ggagggcgta gatggacctg aagtctccag ggacaacagg gcccaggtct
     1201 caggctccta gttgggccca gtggctccag cgtttccaaa cccatccatc cccagaggtt
     1261 cttcccatct ctccaggctg atgtgtggga actcgaggaa ataaatctcc agtgggagac
     1321 ggaggggtgg ccagggaaac ggggcgctgc aggaataaag acgagccagc acagccagct
     1381 catgtgtaac ggctttgtgg agctgtcaag gcctggtctc tgggagagag gcacagggag
     1441 gccagacaag gaaggggtga cctggaggga cagatccagg ggctaaagtc ctgataaggc
     1501 aagagagtgc cggccccctc ttgccctatc aggacctcca ctgccacata gaggccatga
     1561 ttgaccctta gacaaagggc tggtgtccaa tcccagcccc cagccccaga actccaggga
     1621 atgaatgggc agagagcagg aatgtgggac atctgtgttc aagggaagga ctccaggagt
     1681 ctgctgggaa tgaggcctag taggaaatga ggtggccctt gagggtacag aacaggttca
     1741 ttcttcgcca aattcccagc accttgcagg cacttacagc tgagtgagat aatgcctggg
     1801 ttatgaaatc aaaaagttgg aaagcaggtc agaggtcatc tggtacagcc cttccttccc
     1861 tttttttttt ttttttttgt gagacaaggt ctctctctgt tgcccaggct ggagtggcgc
     1921 aaacacagct cactgcagcc tcaacctact gggctcaagc aatcctccag cctcagcctc
     1981 ccaaagtgct gggattacaa gcatgagcca ccccactcag ccctttcctt cctttttaat
     2041 tgatgcataa taattgtaag tattcatcat ggtccaacca accctttctt gacccacctt
     2101 cctagagaga gggtcctctt gcttcagcgg tcagggcccc agacccatgg tctggctcca
     2161 ggtaccacct gcctcatgca ggagttggcg tgcccaggaa gctctgcctc tgggcacagt
     2221 gacctcagtg gggtgagggg agctctcccc atagctgggc tgcggcccaa ccccaccccc
     2281 tcaggctatg ccagggggtg ttgccagggg cacccgggca tcgccagtct agcccactcc
     2341 ttcataaagc cctcgcatcc caggagcgag cagagccaga gcaggttgga gaggagacgc
     2401 atcacctccg ctgctcgcgg ggatctgcga tctaagtaag cttggcattc cggtactgtt
     2461 ggtaaagcca ccatggaaga cgccaaaaac ataaagaaag gcccggcgcc attctatccg
     2521 ctggaagatg gaaccgctgg agagcaactg cataaggcta tgaagagata cgccctggtt
     2581 cctggaacaa ttgcttttac agatgcacat atcgaggtgg acatcactta cgctgagtac
     2641 ttcgaaatgt ccgttcggtt ggcagaagct atgaaacgat atgggctgaa tacaaatcac
     2701 agaatcgtcg tatgcagtga aaactctctt caattcttta tgccggtgtt gggcgcgtta
     2761 tttatcggag ttgcagttgc gcccgcgaac gacatttata atgaacgtga attgctcaac
     2821 agtatgggca tttcgcagcc taccgtggtg ttcgtttcca aaaaggggtt gcaaaaaatt
     2881 ttgaacgtgc aaaaaaagct cccaatcatc caaaaaatta ttatcatgga ttctaaaacg
     2941 gattaccagg gatttcagtc gatgtacacg ttcgtcacat ctcatctacc tcccggtttt
     3001 aatgaatacg attttgtgcc agagtccttc gatagggaca agacaattgc actgatcatg
     3061 aactcctctg gatctactgg tctgcctaaa ggtgtcgctc tgcctcatag aactgcctgc
     3121 gtgagattct cgcatgccag agatcctatt tttggcaatc aaatcattcc ggatactgcg
     3181 attttaagtg ttgttccatt ccatcacggt tttggaatgt ttactacact cggatatttg
     3241 atatgtggat ttcgagtcgt cttaatgtat agatttgaag aagagctgtt tctgaggagc
     3301 cttcaggatt acaagattca aagtgcgctg ctggtgccaa ccctattctc cttcttcgcc
     3361 aaaagcactc tgattgacaa atacgattta tctaatttac acgaaattgc ttctggtggc
     3421 gctcccctct ctaaggaagt cggggaagcg gttgccaaga ggttccatct gccaggtatc
     3481 aggcaaggat atgggctcac tgagactaca tcagctattc tgattacacc cgagggggat
     3541 gataaaccgg gcgcggtcgg taaagttgtt ccattttttg aagcgaaggt tgtggatctg
     3601 gataccggga aaacgctggg cgttaatcaa agaggcgaac tgtgtgtgag aggtcctatg
     3661 attatgtccg gttatgtaaa caatccggaa gcgaccaacg ccttgattga caaggatgga
     3721 tggctacatt ctggagacat agcttactgg gacgaagacg aacacttctt catcgttgac
     3781 cgcctgaagt ctctgattaa gtacaaaggc tatcaggtgg ctcccgctga attggaatcc
     3841 atcttgctcc aacaccccaa catcttcgac gcaggtgtcg caggtcttcc cgacgatgac
     3901 gccggtgaac ttcccgccgc cgttgttgtt ttggagcacg gaaagacgat gacggaaaaa
     3961 gagatcgtgg attacgtcgc cagtcaagta acaaccgcga aaaagttgcg cggaggagtt
     4021 gtgtttgtgg acgaagtacc gaaaggtctt accggaaaac tcgacgcaag aaaaatcaga
     4081 gagatcctca taaaggccaa gaagggcgga aagatcgccg tgtaattcta gagtcggggc
     4141 ggccggccgc ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact
     4201 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     4261 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     4321 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa
     4381 atcgataagg atccgtcgac cgatgccctt gagagccttc aacccagtca gctccttccg
     4441 gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact
     4501 cgtaggacag gtgccggcag cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4561 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4621 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4681 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4741 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4801 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4861 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4921 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4981 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5041 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5101 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5161 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5221 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5281 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5341 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5401 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5461 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5521 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5581 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5641 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5701 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5761 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5821 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5881 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5941 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6001 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6061 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6121 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6181 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6241 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6301 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6361 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6421 gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa
     6481 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     6541 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     6601 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     6661 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     6721 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     6781 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     6841 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     6901 cgcttacaat ttgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     6961 gggcctcttc gctattacgc cagcccaagc taccatgata agtaagtaat attaaggtac
     7021 gggaggtact tggagc
//
